FNR | Ferredoxin-NADP+-oxidoreductase
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AS15 2909 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Chlamydomonas reinhardtii, Chlorella sorokiniana, Synechococcus PCC 6803
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|Recommended dilution||1: 1000 - 3000 with ECL (WB)|
|Expected | apparent MW||
Arabidopsis thaliana, Chlamydomonas reinhardtii, Chlorella sorokiniana, Synechococcus PCC 6803
|Predicted reactivity||Higher plants|
|Not reactive in|
For detection in Chlorella sorokiniana, Synechococcus PCC 6803 higher load per well needs to be applied.
This antibody recognizes all Arabidopsis thaliana FNR isoforms.
To be added when available, antibody released in March 2016.
10 ng of recombinant Chlamydomonas reinhardtii FNR (1), thylakoid preparation (1 µg chl) from Chlamydomonas reinhardtii cc 124 (2), thylakoid preparation (1 µg chl) of Chlorella sorokiniana (3), thylakoid preparation (1 µg chl) of Synechococcus PCC 6803 (4) incubated with urea based sample buffer over night at 25°C were separated on Bolt 4-12 % Bis-Tris Plus gels (Novex, Life Tech) and blotted 1h to nitrocellulose using iBlot Gel Transfer Stacks Nitrocellulose, Mini (Novex, LifeTech). Blots were blocked with iBind (LifeTech) with iBind solution kit (Novex, LifeTech). Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. There is no washing steps using this set up. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602 ) diluted to 1:40 000 in for 1h at RT with agitation. The blot was washed as above and developed for with EZ-ECL (Biological Industries, Israel). Exposure time was 10 min.
Courtesy of M.Sc. Pini Marcu, Tel Aviv University, Israel
Proteins were isolated from wt Arabidopsis thaliana and fnr1 plants with 3X LB (6 M urea, 12% SDS, 30% glycerol, 100 mM DTT, 150 mM Tris pH7.0, 0.8% Comassie G-250). 10 µg of total proteins from leaves or roots were loaded into each lane and separated on 12% SDS-PAGE, and then blotted overnight onto PVDF membrane. Membranes were blocked with milk powder for 2 h and then incubated in the primary antibody solution overnight, which was then decanted and the membrane was washed 3 times for 5 min in TBST. Membrane was incubated at RT for 1 hour in 1:10 000 goat anti-Rabbit secondary antibody from Agrisera, followed by washing steps as above. Membrane was developed for 2 min with Super Signal ECL reagents from Pierce according to the manufacturer’s instructions and recorded using FujiFilm CCD camera with 10 s increment time for around 190 s.
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